Abstract 1091: TWIST1 inhibition overcomes resistance to tyrosine kinase inhibitors in MET driven non-small cell lung cancer
Authors: Vinod Kumar, Zachary A. Yochum, Princey Devadassan, Eric Huang, Ethan Miller, Vasavi Ayyala, Laura P. Stabile, Timothy F. Burns, Purva H. Rumde
Journal: Cancer Research (2022)

Abstract Background: The mesenchymal-epidermal transition (cMET/MET) tyrosine kinase receptor and its ligand, the hepatocyte growth factor (HGF) are overexpressed in a large percentage of non-small cell lung cancers (NSCLCs) and MET mutation and/or amplification leads to oncogene addiction in small subset of NSCLC. In addition, MET amplification is an established mechanism of resistance to EGFR and other oncogenic targeted TKIs. Several MET-inhibitors have been developed and two MET TKIs have been approved for MET exon 14 skipping mutant NSCLC and have shown activity against MET amplified NSCLC. However, long-term efficacy of MET TKIs is limited as acquired resistance is inevitable. HGF overexpression has been identified as one of the mechanisms of resistance to MET TKIs in MET-altered NSCLC, but the mechanism(s) by which the HGF-MET pathway causes resistance are poorly understood. Here, we investigated the requirement of the EMT-transcription factor, TWIST1 in HGF-mediated resistance to MET TKIs and the role of TWIST1 in de-novo and acquired resistance to MET TKIs. Methods: TWIST1 expression was measured in wild type and TWIST1 over expressing cell lines in presence and absence of HGF. TWIST1 was inhibited with shRNA and harmine. Apoptosis was assessed via immunoblotting and cleaved caspase 3 staining. We utilized MET altered (mutant/amplified) NSCLC cell lines, patient derived xenografts and a novel transgenic mouse model of Hgf, Twist1 overexpressing lung cancer to evaluate TWIST1 as a driver of MET TKI resistance. Results: We found that HGF induced TWIST1 expression and MET TKI treatment decreased TWIST1 expression through a post-translational mechanism. Re-expression of TWIST1 led to MET TKI resistance in MET amplified or MET mutant cell lines. Conversely, genetic and pharmacological inhibition of TWIST1 sensitizes to MET inhibition and overcame HGF-mediated MET TKI resistance and MET amplification mediated EGFR TKI resistance in vitro and in vivo. Furthermore, the role of TWIST1 in HGF/MET lung tumorigenesis was evaluated both in human NSCLC xenografts and an Hgf-driven NSCLC mouse model. Genetic inhibition of TWIST1 in MET mutant or amplified cell lines prevented tumor growth in vivo. Furthermore, TWIST cooperated with Hgf in a CCSP-Hgf (CH) mice model that constitutively overexpresses Hgf in the lung and develops NSCLC after treatment with the tobacco carcinogen, 4-(methylnitrosoamino)-1-(3-pyridyl)-1-butanone (NNK) was utilized. We demonstrated that the Twist1 overexpressing CHT (CCSP-rtTA/CCSP-Hgf/Twist1-tetO-luc) mice developed significantly larger and more aggressive tumors as compared to CH and CCSP-rtTA/Twist1-tetO-luc (CT) mice and that continued TWIST1 expression was required for these tumors. Conclusions: Our findings suggest that targeting TWIST1 may be an effective therapeutic strategy to overcome HGF-MET-driven resistance in MET-driven NSCLC. Citation Format: Vinod Kumar, Zachary A. Yochum, Princey Devadassan, Eric Huang, Ethan Miller, Vasavi Ayyala, Laura P. Stabile, Timothy F. Burns, Purva H. Rumde. TWIST1 inhibition overcomes resistance to tyrosine kinase inhibitors in MET driven non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1091.